Anatomy Commons^™

Nadph Oxidase As A Therapeutic Target In Alzheimer's Disease, Michelle L. Block

Anatomy and Neurobiology Publications

At present, available treatments for Alzheimer's disease (AD) are largely unable to halt disease progression. Microglia, the resident macrophages in the brain, are strongly implicated in the pathology and progressively degenerative nature of AD. Specifically, microglia are activated in response to both β amyloid (Aβ) and neuronal damage, and can become a chronic source of neurotoxic cytokines and reactive oxygen species (ROS). NADPH oxidase is a multi-subunit enzyme complex responsible for the production of both extracellular and intracellular ROS by microglia. Importantly, NADPH oxidase expression is upregulated in AD and is an essential component of microglia-mediated Aβ neurotoxicity. Activation of …

A Synaptic Nidogen: Developmental Regulation And Role Of Nidogen-2 At The Neuromuscular Junction, Michael A. Fox, Matthew Sp Ho, Neil Smyth, Joshua R. Sanes

Anatomy and Neurobiology Publications

Background

The skeletal neuromuscular junction is a useful model for elucidating mechanisms that regulate synaptogenesis. Developmentally important intercellular interactions at the neuromuscular junction are mediated by the synaptic portion of a basal lamina that completely ensheaths each muscle fiber. Basal laminas in general are composed of four main types of glycosylated proteins: laminins, collagens IV, heparan sulfate proteoglycans and nidogens (entactins). The portion of the muscle fiber basal lamina that passes between the motor nerve terminal and postsynaptic membrane has been shown to bear distinct isoforms of the first three of these. For laminins and collagens IV, the proteins are …

Arabidopsis Mrna Polyadenylation Machinery: Comprehensive Analysis Of Protein-Protein Interactions And Gene Expression Profiling, Arthur G. Hunt, Ruqiang Xu, Balasubrahmanyam Addepalli, Suryadevara Rao, Kevin P. Forbes, Lisa R. Meeks, Denghui Xing, Min Mo, Hongwei Zhao, Amrita Bandyopadhyay, Lavanya Dampanaboina, Amanda Marion, Carol Von Lanken, Qingshun Quinn Li

Anatomy and Neurobiology Publications

Background

The polyadenylation of mRNA is one of the critical processing steps during expression of almost all eukaryotic genes. It is tightly integrated with transcription, particularly its termination, as well as other RNA processing events, i.e. capping and splicing. The poly(A) tail protects the mRNA from unregulated degradation, and it is required for nuclear export and translation initiation. In recent years, it has been demonstrated that the polyadenylation process is also involved in the regulation of gene expression. The polyadenylation process requires two components, the cis-elements on the mRNA and a group of protein factors that recognize the cis …

Protocol: Streamline Cloning Of Genes Into Binary Vectors In Agrobacterium Via The Gateway® Topo Vector System, Ruqiang Xu, Qingshun Quinn Li

Anatomy and Neurobiology Publications

Background

In plant functional genomic studies, gene cloning into binary vectors for plant transformation is a routine procedure. Traditionally, gene cloning has relied on restriction enzyme digestion and ligation. In recent years, however, Gateway^® cloning technology (Invitrogen Co.) has developed a fast and reliable alternative cloning methodology which uses a phage recombination strategy. While many Gateway- compatible vectors are available, we frequently encounter problems in which antibiotic resistance genes for bacterial selection are the same between recombinant vectors. Under these conditions, it is difficult, if not sometimes impossible, to use antibiotic resistance in selecting the desired transformants. We have, …

The Effects Of Atomoxetine On Cognitive Performace And Neuroplasticity After Traumatic Brain Injury, Wendy Reid

Theses and Dissertations

Catecholaminergic neurotransmission is regionally altered following injury, and drugs aimed at these systems offer promising avenues for post-TBI pharmacotherapies. Atomoxetine is a selective norepinephrine transporter (NET) inhibitor currently indicated for treatment of attention-deficit hyperactivity disorder (ADHD). The studies in this dissertation were designed to test the efficacy of atomoxetine for treating cognitive deficits following experimental TBI and the potential mechanism for any beneficial effect. The first part of the study focused on behavioral recovery following atomoxetine treatment. Several important questions of dose, therapeutic window, and duration of treatment were addressed in these studies. Sprague-Dawley rats were subjected to lateral fluid-percussion …

The Role Of Inhibitory Interneurons In A Model Od Developmental Epilepsy, Patrick James Wolfgang

Theses and Dissertations

Epilepsy, defined by recurrent seizures, is the one of the most prevalent neurological disorders worldwide (World Health Organization 2007). While many forms of epilepsy are well-controlled by anti-epileptogenic medications, a significant portion of patients have intractable, i.e. untreatable, seizures. The etiology of these seizures is varied, but a significant cause, particularly for patients with intractable epilepsy is developmental malformation. In these cases, an error or interruption during the development of the neocortex produces a structural alteration. Such patients may have other neurological problems, but seizures are the most common symptom. The neuronal mechanisms that link malformation and cortical hyperexcitability are …

Analysis Of Mouse Eklf/Klf2 E9.5 Double Knockout: Yolk Sac Morphology And Embryonic Erythroid Maturation, Tina Kathy Lung

Theses and Dissertations

Krüppel-like factors (KLFs) are a family of transcription factors with 3 Cys2/His2 zinc fingers that regulate cell differentiation and developmental processes. EKLF is involved in primitive and definitive erythropoiesis; KLF2 is implicated in the development of primitive erythroid and endothelial cells of the vasculature. Using light and electron microscopy, the yolk sacs and dorsal aortae from EKLF/KLF2 double knockout (KO) E9.5 (embryonic day 9.5) were examined to determine whether these KLFs have compensatory functions in morphology of blood cells and vessels. EKLF/KLF2 double KO E9.5 erythroid, endothelial, and mesothelial cells had more severely abnormal morphology than WT and KLF2-/-. Flow …

Assessing Epidermal Growth Factor Expression In The Rodent Hippocampus Following Traumatic Brain Injury, Janice Mabutas Daus

Theses and Dissertations

Hippocampal neurons are vulnerable to injury, as indicated by the prevalence of learning and memory deficits following traumatic brain injury. Research indicates that proliferation of neural precursor cells increases following brain injury, which implies that there is an endogenous response in the hippocampus to replenish neurons and restore cognitive function. Studies show that mitogenic growth factors may drive this proliferative response; one of which is epidermal growth factor. Because adults and the elderly manifest the most enduring deficits following TBI, it is critical to investigate how EGF expression following injury may relate to injury-induced cell proliferation and the degree of …

The Effect Of Silencing The Wilms' Tumor 1 Gene On The Radiation Sensitivity Of Glioblastoma Cells, Dana C. Chan

Theses and Dissertations

Glioblastomas are among the most devastating of human cancers with a median survival of only 9-12 months. This type of brain tumor is incurable, largely due its remarkable proliferative capacity and resistance to current treatments. High levels of the Wilms' Tumor 1 (WTI) gene have been identified in glioblastomas, suggesting an oncogenic function. Moreover, known WT1 target genes have been implicated in resistance to radiation. To determine the role of WT1 in radiation resistance, two glioblastoma cell lines expressing WT1 were treated with siRNAs to silence this gene. Confirmation of WT1 knockdown was achieved through real-time PCR and Western blot. …

Diffuse Brain Injury Triggers Ultra-Rapid Perisomatic Traumatic Axonal Injury, Wallerian Change, And Non-Specific Inflammatory Responses, Brian Joseph Kelley

Theses and Dissertations

A significant component of diffuse brain injury (DBI) is diffuse axonal injury (DAI) which is responsible for the morbidity and mortality associated with this condition. DAI and its experimental counterpart traumatic axonal injury (TAI) result in scattered microscopic pathology characterized by focal impairment of axonal transport leading to progressive swelling and delayed axotomy. DBI-mediated perisomatic axotomy does not result in acute neuronal death suggesting that delayed axotomy was responsible for this unanticipated response. To evaluate this hypothesis, we examined the spatiotemporal progression of DBI-mediated perisomatic TAI. LM / TEM identified impaired axonal transport within 15 - 30 min post-injury. Perisomatic …

The Expression And Function Of Wilms' Tumor 1 In Malignant Glioma, Aaron J. Clark

Theses and Dissertations

The Wilms' tumor 1 gene is overexpressed in many types of cancer and is associated with poor prognosis and resistance to anti-cancer therapies. In vitro studies in non-glioma cells types have demonstrated that WTl plays a role in increased proliferation, resistance to apoptosis, and increased cellular invasion. We aimed to thoroughly characterize the expression pattern of Wilms' tumor 1 in human malignant glioma and discern its function in this complex disease process. We screened a large sample of established human malignant glioma cell lines and glioma tissue specimens of all grades for WT1 expression. The majority of cell lines and …

A Model For Studying Vasogenic Brain Edema, Anshu Shukla

Theses and Dissertations

Convection-enhanced delivery (CED) is a proven method for targeted drug delivery to the brain that circumvents the blood-brain barrier (BBB). Little study has been conducted in understanding CED in pathological brain states. This is of importance when dealing with chemotherapeutic agent delivery to brain tumors, where vasogenic edema (VE) exists. The current study aims to characterize a model of VE suitable for studying CED.VE was produced in the right hemisphere of the rat brain using multiple infusions of hyperosmotic mannitol (0.25mL/kg/s over 30 seconds) delivered through the right internal carotid artery. Magnetic resonance imaging (MRI) revealed consistent edema formation and …

Characterization And Functional Analysis Of A Newly Identified Human Mt5-Mmp Transcript Variant Isolated From Multipotent Nt2 Cells, Heather Hamilton Ross

Theses and Dissertations

Membrane-type 5 matrix metalloproteinase (MT5-MMP) is unique among MMP family members as it is predominately expressed in the CNS. Its expression is ubiquitous during brain development and restricted to regions of neurogenesis and neuroplasticity in the adult. MT5-MMP is a mediator of pericellular proteolysis and is thought to have a functional impact on neurite outgrowth. The studies presented in this work were designed to examine MT5-MMP expression in cultured NT2 cells, a model of newogenesis and neuronal differentiation, and in adult neurogenic brain regions. We further sought to overexpress MT5-MMP and test the hypothesis that it plays a role in …

Cross-Modal Projections From Auditory To Visual Cortices In The Ferret, Meng Y. Wang

Theses and Dissertations

Recent studies have shown that neuronal connections occur between primary auditory and visual cortices of the primate (Falchier et al., 2002; Rockland and Ojima, 2003), and it has been suggested that these projections are involved in multisensory processing in these lower-level, core areas of cortex. The present study was conducted to determine if similar connections occur in other higher mammals such as carnivores (ferrets; Mustela putorius). Large injections of sensitive neuroanatomical tracer were placed within the core areas of auditory cortex in 3 ferrets. After transport and processing, labeled axon terminals were found not in primary visual cortex, but in …

Combinatorial Modulation Of Multiple Signaling Pathways To Gain Therapeutic Response In Breast And Prostate Cell Carcinomas, William Tressel Hawkins Ii

Theses and Dissertations

Our laboratory is primarily interested in novel pharmacological intervention of cell proliferation and survival pathways expressed in various types of cancer. These cyto-protective pathways can be activated in response to growth factor stimulation, toxic insult and radiation. In our studies, we utilized novel drug combinations with and without radiation to enhance breast & prostate tumor cell death both in vitro and in vivo. Previous studies from our group have shown that UCN-01 and MEK1/2 inhibitors interact to cause tumor cell death in transformed cell lines in vitro. We extended this observation to an in vivo animal model system using the …

The Role Of Sulfatide In Alzheimer's Disease, Charles Britton Beasley Jr.

Theses and Dissertations

Alzheimer's disease (AD) is characterized by the accumulation of amyloid beta plaques, neurofibrillary tangles (NFT) and loss of cortical neurons that control memory and cognition. The cause of NFTs and Aβ plaques is not clear, though it is known that they are formed by enzymes which are preferentially sequestered to membrane domains called lipid rafts. Sulfatide (ST) is a glycosphingolipid that is essential for the proper structure and function of lipid rafts. In mice that lack ST, membrane domains that are normally maintained by adhesive contacts and functional lipid rafts are improperly formed and are unstable. In these ST null …

The Influence Of A Single Nucleotide Polymorphism In The Matrix Metalloproteinase-1 Promoter On Glioma Biology, Jessica Mccready

Theses and Dissertations

Glioblastomas are an incurable type of brain tumor with a mean survival time of 9-12 months following diagnosis. One of the reasons for this poor prognosis is the ability of tumor cells to invade the surrounding normal brain tissue. Enzymes responsible for this invasive nature include the matrix metalloproteinase family. MMP-1 is a member of this family which has been well studied in many types of invasive tumors, with gliomas being an exception. We studied a single nucleotide polymorphism (SNP) in the MMP-1 promoter that may influence glioma biology. This SNP consists of the presence (2G) or absence (1G) of …

Molecular Localization Of Hypoxia Inducible Factor-1-Alpha In Post-Ischemic Myocardium Following In Vivo Prolyl-4 Hydroxylase-2 Gene Silencing, Julia Antoinette Messina

Theses and Dissertations

Administration of small interfering RNA (siRNA) specific for prolyl-4 hydroxylase-2 (PHD2) results in PHD2 inhibition, Hypoxia Inducible Factor-I (HIF-1) activation, and cardioprotection versus Ischemia Reperfusion (IR). This study observes the effects of siRNA-mediated PHD2 inhibition on the distribution of cardioprotective proteins by immunofluorescence and basic histology. Fifteen mice were divided into 5 groups: PHD2 Control, Non-Targeting scramble (NTS) Control, IR Control, PHD2 IR, and NTS IR. Histologically, tissue damage was reduced dramatically in the PHD2 IR group compared to the NTS IR and IR control groups. From confocal images, total fluorescent pixels and intensities were quantified. The PHD2 IR group …

Identification And Characterization Of The Expression Profile Of Oligodendrocyte-Derived And Associated Proteins Via Unilateral X-Irradation Of The Rat Optic Nerve, Nicholas Greco

Theses and Dissertations

Recent studies examining cell-cell interactions during CNS development and following disease or trauma have highlighted our limited understanding of the in vivo functions of the myelinating cell of the CNS, the oligodendrocyte. With this in mind, our laboratory has developed techniques by which a profile of proteins derived from or regulated by oligodendrocytes can be elucidated. Specifically, we have demonstrated that oligodendrocytes can be selectively eliminated from one optic nerve of a rat by treating the animal with a unilateral exposure of X-irradiation at the time of birth. Consequently, this approach allowed us to experimentally create, within the same animal, …

Status Epilepticus Results In A Duration-Dependent Increased Protein Kinase A Activity In The Rat Pilocarpine Model, James M. Bracey

Theses and Dissertations

This study was conducted to characterize cellular changes occurring during the progression of status epilepticus (SE) that could lead to the maintenance of increased membrane excitability. SE was induced by injection of pilocarpine after which rats were monitored both electrographically and behaviorally. After various lengths of time in SE, specific brain regions were isolated for biochemical study. SE resulted in an early maintenance of PKA activity in both cortical homogenate and crude synaptoplasmic membrane (crude SPM) fractions. At subsequent stages of SE there was a significant increase in PKA activity in both homogenate and crude SPM fractions. Wester blot analysis …

The Effect Of Traumatic Brain Injury On Expression Levels Of Ankyrin-G In The Corpus Callosum And Cerebral Cortex, Andrew S. Vanderveer

Theses and Dissertations

The ankyrins comprise a family of proteins serving as components of the membrane cytoskeleton, and participate in a diverse set of associations with multiple binding partners including the cytoplasmic domains of transporters, ion channels, some classes of receptors, and cell adhesion proteins. Moreover, evidence is accumulating that ankyrin participates in defining functionally distinct subcellular regions. The complex functional and structural roles of ankyrins indicate they are likely to play essential roles in the pathology of traumatic axonal injury. The current study examined changes in ankyrin-G expression following a moderate central fluid percussion injury administered to adult rats. At 1d, 3d, …

Mechanisms Underlying Protection Against Rt-2 Gliomagenesis In Rat Brain Utilizing Primary And Secondary Vaccination, Andrea M. Lister

Theses and Dissertations

Primary brain tumor affects some 18,000 adults in the United States each year (Silverberg et al., 1990; Merchant et al., 1997) and over 30% are high-grade anaplastic astrocytoma or glioblastoma multiforme (GBM) (Parney et al., 1997). According to Kruse et al., 1989, the treatment of patients with recurrent or persistent high-grade gliomas represents a major therapeutic challenge. The use of conventional therapy consisting of surgery, followed by radiation therapy and chemotherapy for gliomas, has been relatively ineffective (Jaecle et al., 1994) despite the fact that these therapies are cytoreductive in nature (Black, 1991). Most malignancies will recur locally but may …

The Pineal Gland, Via Melatonin, Protects Dna, Coordinates The Endocrine System With The Immune System And Controls The Timing Of Reproduction, Craig L. Anthony

Theses and Dissertations

The pineal gland secretes the hormone melatonin (n-acetyl-5-methoxytryptamine) with a circadian rhythm. This secretion's rhythm becomes disrupted with age. When melatonin secretion is decreased with advanced age, the immune, endocrine and reproductive systems fail to function optimally. Melatonin possesses lipophilic and anti-oxidant properties, providing it with access to nuclei. Melatonin protects the DNA, preventing cancerous mutation. Hippocampal degeneration and age increase and prolong the adrenocortical responses to stress. Melatonin supplementation reduces the prolonged exposure to harmful hormones.

Study Of P53 Gain Of Function Mutations In P53-Null Astrocytes, Sang H. Choi

Theses and Dissertations

A number of recent studies suggest that expression of mutant p53 with mutations in certain codons show a gain of function and some of the characteristics of an oncoprotein. In order to study gain of function mutation and eliminate the potential of a dominant negative interaction with endogenous wild type p53 protein, p53 knockout mouse astrocytes were used. A retrovirus system was used to introduce mutant p53 genes into these p53-null astrocytes. Immunohistochemical staining and western blot experiments showed the expression of mutant p53 protein in these cells after infection with the mutant p53 retroviruses. Cell growth experiment did not …

Excisional Wound Healing And The Role Of Homeobox Genes, David Allen Lanning

Theses and Dissertations

The biochemical and physiologic mechanisms responsible for excisional wound contraction have been extensively studied but are not completely understood. Furthermore, the genes that regulate these processes are unknown. As a result, there are limited treatment options available for patients with diseases or conditions characterized by abnormal tissue contraction. In an attempt to improve our understanding of these mechanisms and identify some of the putative regulatory genes, we studied excisional wound healing in the mid-gestational fetal rabbit. These wounds normally do not contract nor are they associated with inflammation, fibrosis, or scar formation. However, we have been able to induce contraction, …

Gene Expression Following Traumatic Brain Injury, Rajiv Malhotra

Theses and Dissertations

The pathology which results from traumatic brain injury (TBI) have long been believed to be immediate and irreversible. However, recently it has been shown that, although the primary effects are virtually unavoidable, the secondary effects manifest themselves through biochemical processes set in motion at the time of the injury. These events are frequently mediated through the process of excitotoxicity, which results from a widespread release of excitatory neurotransmitters. These neurotransmitters go on to activate both ionotropic and metabotropic receptors. The signal transduction initiated through these receptor populations gives rise to changes in gene expression.

One result of this release of …

Tumor Immunity Following Adenovirus Mediated Herpes Simplex Thymidine Kinase Gene Transfer To Experimental Rat Gliomas, Maulik Raj Shah

Theses and Dissertations

Previous studies have determined adenovirus mediated herpes simplex thymidine kinase gene transfer (AV-TK) to be effective for the treatment of experimental gliomas. In this study we report three distinct phenomenon. First, animals with complete regression of subcutaneous tumors upon intratumoral injections of AV-TK with concomitant Ganciclovir® (GCV) administration developed tumor immunity. These animals had the ability to reject a subsequent inoculum of lethal doses of tumor cells. This tumor immunity was long standing and protective as far as 6 months from the time of initial tumor ablation. Of interest, adoptive transfer of splenocytes from AV-TK treated-tumor ablated animals to naive …

Computer Modeling Of Anatomical Structure: A Representative Example Of Modeling The Inguinal Canal, Jason Michael Highsmith

Theses and Dissertations

As computers become an increasingly important part of medical education, a proper understanding of the techniques and applications of computer aided modeling is vital. An initial overview of medical imaging and the techniques of computer modeling is presented. Construction of three-dimensional models of anatomical structures is then discussed in great detail with specific focus on modeling structures like the inguinal canal. The inguinal canal is one region where computer modeling efforts should be directed because it presents a special challenge. Understanding the walls, borders and layering of the inguinal canal is especially difficult but vital to accurate clinical diagnoses of …

Nmda Receptor-Mediated Synaptic Plasticity In Developing Mammalian Visual Pathways, Gregory S. Perens

Theses and Dissertations

Precise connections in many mammalian nervous systems require a great deal of remodeling during development. In the visual system, many excess synapses are originally formed in the lateral geniculate nucleus and striate cortex. Only the correct set of axon terminals are retained during normal development, while imprecise ones withdraw. The mechanism by which only correct axons are retained requires neural activity, and may be regulated by specific receptors at synapses.

The transmission of neural signals at these synapses is carried out in part by the glutamate-activated NMDA receptor. It is hypothesized that NMDA receptor activation plays a crucial role in …

The Impact Of The Tau Mutation On Reproductive Function In The Golden Hamster, Eliza Leake Thomasson

Theses and Dissertations

The tau mutation affects the circadian system of the golden hamster primarily by decreasing the period of the activity cycle from 24 to 20 hours. To study the effect of this mutation on reproductive function in the golden hamster, 3 experiments have been designed. In the first experiment, wild-type and tau female hamsters will be maintained in conditions of 14:10 LD and 11.7:8.3 LD, respectively. Blood samples will be taken and analyzed by radioimmunoassay (RIA) to determine the timing of the proestrus LH surge. In the second experiment, wild-type and tau males will be transferred to shortened photoperiods of 10:14 …